Herpes simplex virus type 1 immediate-early protein Vmw110 reactivates latent herpes simplex virus type 2 in an in vitro latency system.

Reactivation of latent herpes simplex virus type 2 (HSV-2) by the immediate-early protein Vmw110 was studied by using an in vitro latency system. Adenovirus recombinants that express Vmw110 reactivated latent HSV-2. An HSV-1 mutant possessing a deletion in a carboxy-terminal region of Vmw110 reactivated latent HSV-2, whereas mutant FXE, which has a deletion in the second exon, did not. Therefore, Vmw110 alone is required to reactivate latent HSV-2 in vitro, and the region of Vmw110 defined by the deletion in FXE is important for this process.     

Bactericidal activity of peroxynitrite.

Peroxynitrite is a strong oxidant formed by macrophages and potentially by other cells that produce nitric oxide and superoxide. Peroxynitrite was highly bactericidal, killing Escherichia coli in direct proportion to its concentration with an LD50 of 250 microM at 37 degrees C in potassium phosphate, pH 7.4. The apparent bactericidal activity of a given concentration peroxynitrite at acidic pH was less than that at neutral and alkaline pH. However, after taking the rapid pH-dependent decomposition of peroxynitrite into account, the rate of the killing was not significantly different at pH 5 compared to pH 7.4. Metal chelators did not decrease peroxynitrite-mediated killing, indicating that exogenous transition metals were not required for toxicity. The hydroxyl radical scavengers mannitol, ethanol, and benzoate did not significantly affect toxicity while dimethyl sulfoxide enhanced peroxynitrite-mediated killing. Dimethyl sulfoxide is a more efficient hydroxyl radical scavenger than the other three scavengers and increased the formation of nitrogen dioxide from peroxynitrite. In the presence of 100 mM dimethyl sulfoxide, 60.0 +/- 0.3 microM nitrogen dioxide was formed from 250 microM peroxynitrite as compared to 2.0 +/- 0.1 microM in buffer alone. Thus, formation of nitrogen dioxide may have enhanced the toxicity of peroxynitrite decomposing in the presence of dimethyl sulfoxide.     

Time-Dependent Increase in Protein Phosphorylation Following One-Trial Enhancement in Hermissenda

Abstract: One-trial conditioning of the nudibranch mollusk Hermissenda produces short- and long-term changes in excitability (enhancement) of identified sensory neurons. To investigate the biochemical mechanisms underlying this example of plasticity, we have examined changes in protein phosphorylation at different times following the in vitro conditioning trial. Changes in the incorporation of ³²PO₄ into proteins were determined using two-dimensional polyacrylamide gel electrophoresis, autoradiography, and densitometry. Conditioning resulted in increases in levels of several phosphoproteins, five of which, ranging in apparent molecular mass from 22 to 55 kDa, were chosen for analysis. The increased phosphorylation of the 46- and 55-kDa phosphoproteins detected 2 h postconditioning was significantly greater than the level of phosphorylation detected in an unpaired control group, indicating that long-term enhancement is pairing specific. Statistically significant increases in phosphorylation as compared with the control group that received only light were detected immediately after conditioning (5 min) for the 55-, 46-, and 22-kDa phosphoproteins, at 1 h for the 55- and 46-kDa phosphoproteins, and at 2 h for the 55-, 46-, and 22-kDa phosphoproteins. The 46- and 55-kDa phosphoproteins are putative structural proteins, and the 22-kDa phosphoprotein is proposed to be a protein kinase C substrate previously identified in Hermissenda following multitrial classical conditioning. Time-dependent increases in protein phosphorylation may contribute to the induction and maintenance of different memory stages expressed in sensory neurons after one-trial conditioning.     

Regeneration of fertile transgenic indica (group 1) rice plants following microprojectile transformation of embryogenic suspension culture cells

Regenerable embryogenic suspensions of elite Indica (group 1) rice varieties IR24, IR64, IR72 and an advanced Indica rice breeding line IR57311-95-2-3 were established within 6–8 weeks from 3–4 week old calli derived from mature seeds. Transgenic rice plants were obtained by introducing a plasmid carrying genes encoding hygromycin phosphotransferase (hph, conferring resistance to hygromycin B) and ß-glucuronidase (uidA), both driven by the CaMV 35S promoter, via particle bombardment of embryogenic suspensions. The effect of osmotic conditioning on transformation was evaluated. Regenerated plants were resistant to hygromycin B and expressed the uidA (GUS) gene. The growth of mother plants (R₀) was normal and seeds were produced. Southern blot analysis of R₀ and R₁ plants showed that hygromycin resistant plants contained intact hph genes that were inherited in a Mendelian fashion. A protocol for a simple, efficient, repeatable, genotype- and environment-independent Indica rice transformation system is described.Abbreviations 2,4-D 2,4-dichlorophenoxy acetic acid - NAA -naphthalene acetic acid - kb kilobase - GUS ß-glucuronidase - hph hygromycin B phosphotransferase     

The effects of mycorrhizal roots on litter decomposition, soil biota, and nutrients in a spodosolic soil

We studied the effects of mycorrhizal pitch pine (Pinus rigida) roots on litter decomposition, microbial biomass, nematode abundance and inorganic nutrients in the E horizon material of a spodosolic soil, using field microcosms created in a regenerating pitch pine stand in the New Jersey Pinelands. Pine roots stimulated litter decomposition by 18.7% by the end of the 29 month study. Both mass loss and N and P release from the litter were always higher in the presence of roots than in their absence. Nutrient concentrations in decomposing litter were similar, however, in the presence and absence of roots, which suggests that the roots present in the with-root treatment did not withdraw nutrients directly from the litter. The soil was slightly drier in the presence of roots, but there was no discernible effect on soil microbial biomass. The effects of roots on soil extractable inorganic nutrients were inconsistent. Roots, however, were consistently associated with higher numbers of soil nematodes. These results suggest that, in soils with low total C and N contents, roots stimulate greater activity of the soil biota, which contribute, in turn, to faster litter decomposition and nutrient release.Contribution No. 95-22 from the Institute of Marine and Coastal Sciences.Contribution No. 95-22 from the Institute of Marine and Coastal Sciences.     

Bi（Ⅲ）与金属硫蛋白作用性质研究

 Ｂｉ（Ⅲ）与金属硫蛋白作用性质研究张保林,黄辉,朱凌燕,岳晟,唐雯霞（南京大学配位化学研究所,配位化学国家重点实验室,南京２１００９３）如何降低顺铂或其它抗癌铂的毒性,一直是癌症化疗中的重要课题之一,最近研究发现预先给大鼠或肺癌病人服用铋盐,可以极大...     

Intracellular transport of newly synthesized varicella-zoster virus: final envelopment in the trans-Golgi network.

The maturation and envelopment of varicella-zoster virus (VZV) was studied in infected human embryonic lung fibroblasts. Transmission electron microscopy confirmed that nucleocapsids acquire an envelope from the inner nuclear membrane as they enter the perinuclear-cisterna-rough endoplasmic reticulum (RER). Tegument is not detectable in these virions; moreover, in contrast to the mature VZV envelope, the envelope of VZV in the RER is not radioautographically labeled in pulse-chase experiments with [3H]mannose, and it lacks gpI immunoreactivity and complex oligosaccharides. This primary envelope fuses with the RER membrane (detected in cells incubated at 20 degrees C), thereby releasing nucleocapsids to the cytosol. Viral glycoproteins, traced by transmission electron microscopy radioautography in pulse-chase experiments with [3H]mannose, are transported to the trans-Golgi network (TGN) by a pathway that runs from the RER through an intermediate compartment and the Golgi stack. At later chase intervals, [3H]mannose labeling becomes associated with enveloped virions in post-Golgi locations (prelysosomes and plasma membrane). Nucleocapsids appear to be enveloped by wrapping in specialized cisternae, identified as the TGN with specific markers. Tegument-like material adheres to the cytosolic face of the concave surface of TGN sacs; nucleocapsids adhere to this protein, which is thus trapped between the nucleocapsid and the TGN-derived membrane that wraps around it. Experiments with brefeldin A suggest that tegument may bind to the cytosolic tails of viral glycoproteins. Fusion and fission convert the TGN-derived wrapping sacs into an inner enveloped virion and an outer transport vesicle that carries newly enveloped virions to cytoplasmic vacuoles. These vacuoles are acidic and were identified as prelysosomes. It is postulated that secreted virions are partially degraded by their exposure to the prelysosomal internal milieu and rendered noninfectious. This process explains the cell-associated nature of VZV in vitro; however, the mechanism by which the virus escapes diversion from the secretory pathway to the lysosomal pathway in vivo remains to be determined.     

Effect of selenium and protein deficiency on selenium and glutathione peroxidase in rats

Twenty-four weanling male Wistar rats were divided into four groups fed diets containing adequate or deficient levels of selenium (0.5 ppm [+ Se] or <0.02 ppm [−Se] and protein (15% [+Pro] or 5% [−Pro]), but adequate levels of all other nutrients for 4 wk to determine the effects of Se deficiency and protein deficiency on tissue Se and glutathione peroxidase (GSHPx) activity in rats. Plasma, heart, liver, and kidney Se and GSHPx were significantly lower in Se-deficient groups in relation to Se-sufficient groups. In Se-deficient groups, Se and GSHPx were significantly higher in −Se−Pro rats in heart, liver, and kidney. Data analysis showed that there were significant interaction effects between dietary Se and protein on Se and GSHPx of rats. It is assumed that under the condition of Se deficiency. a low level of protein may decrease Se and GSHPx utilization, increase GSHPx synthesis, and result in Se redistribution. This could account for high levels of Se and GSHPx in the −Se−Pro rats compared to −Se+Pro rats.     

人血红细胞酪氨酸蛋白磷酸酶（PTPP）的研究Ⅱ·胞浆PTPP的分离纯化及其主要性质

人血红细胞胞浆部分经（ＮＨ_４）_２ＳＯ_４沉淀,ＤＥＡＥ－纤维素（ＤＥ５２）柱层析,磷酸纤维素柱层析（Ｐ１１）得到部分纯化的ＰＴＰＰ,产率：５．７％,提纯１０７５倍。以（３２）￣Ｐ－Ｔｙｒ－Ｐｏｌｙ（Ｇ_４：Ｔ）作底物,测得其表征Ｋｍ约为０．５－０．８μｍｏｌ／Ｌ,该酶的最适ｐＨ和最适温度分别为７．０－７．８及３７－４０℃。Ｚｎ￣（２＋）等二价金属离子及Ｎａ_３ＶＯ_４等酸根基团对其活性有明显的抑制作用;ＥＤＴＡ、甘油及ＤＴＴ、巯基乙醇等则对其有强烈激活作用;而氟化物、酒石酸等对ＰＴＰＰ活性基本无影响。此外,某些蛋白质、氨基酸、核苷酸及抗肿瘤药物等对ＰＴＰＰ活性也都有不同程度的影响。特别是一些ＰＫｓ及ＰＰｓ在体外对ＰＴＰＰ活性也具有不同的作用。     

人血红细胞蛋白质酪氨酸磷酸酶（PTPP）的研究Ⅰ·PTPP在红细胞胞浆和膜中的分布及其影响因素

通过测定红细胞胞浆及膜中的ＰＴＰＰ活性,发现人正常血红细胞中胞浆ＰＴＰＰ活性约占红细胞ＰＴＰＰ总活性的７０－８０％,膜中只有约２０－３０％的ＰＴＰＰ活性。很多因素诸如：病变、ＰＨ值、温度、离子强度、细胞贮存时间以及药物等,对ＰＴＰＰ在胞浆与膜中的分布有影响。可以推测：膜上的ＰＴＰＰ能通过某种机制解离下来,进入胞浆;相反的过程,胞浆中的ＰＴＰＰ也能通过某种机制与膜结合。这种偶联与去偶联的具体机制及其生理功能还有待进一步探索。